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Somatic embriogenesis and regeneration of Vitis sp.
41-45.Views:245Somatic embriogenesis and regeneration of Vitis sp.
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Agrobacterium transformation of Rhodiola sp.: current status and limitations
37-42.Views:275The study of secondary metabolites has led to the discovery of new drugs for treating human diseases. However, consistent plant supply can be challenging, leading to the use of plant tissue culture techniques such as hairy root culture. Hairy roots have stable genetics, lateral branching, and can produce secondary metabolites, including alkaloids, flavonoids, and terpenoids. Research on hairy roots as a subject began in the late 19th century, and for the last four decades, hairy roots have been utilized for producing secondary metabolites and recombinant proteins. This article focuses on Rhodiola species - genus of perennial plants that belongs to the family Crassulaceae - and its potential as a source of secondary metabolites using hairy root culture techniques. Rhodiola sp. is widely distributed throughout the Arctic regions of the Northern Hemisphere, with several species having significant medicinal properties. The article discusses the possible use of hairy root cultures for the production of Rhodiola secondary metabolites, including salidroside and rosavins, which have demonstrated significant pharmacological activity in various studies. The use of elicitation and genetic engineering techniques to boost secondary metabolite production in Rhodiola hairy roots is also explored. Overall, the article highlights the potential of Rhodiola hairy root cultures as a valuable source of secondary metabolites with medicinal properties. However, despite some studies Rhodiola hairy root induction and culturing still remains highly unexplored.
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Nutritional and seasonal requirements for callus growth in Taxus baccata
111-114.Views:180Callus cultures derived from young stems of two varieties of Taxus baccata cv. aureovariegata (genotype I) and Taxus baccata L. (genotype II, III) were induced. Gamborg's B5 medium was supplemented with different concentrations of auxin (2,4-D) in combination with cytokinins (kinetin or topolin) and with a phenolic-binding compound (PVP) to prevent callus darkening and growth inhibition. Stem explants displayed different responses to in vitro culture depending on plant genotype and on the season. Genetic variability was observed in the growth rate of calli initiated from all three genotypes of the same Taxus species. We found the best growth of callus cultures originated from the genotype ill in defined media. After the first subculture the majority of the cream-coloured primary callus turned brown and ceased its growth. However, the long-term culture was initiated.
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Bean tissue culture and genetic transformation with Agrobacterium
32-35.Views:270In this paper we report the establishment methods of a rapidly growing callus culture of Phaseolus vulgaris bean as well as the conditions required for a high level of transient gene expression using Agrobacterium-mediated transformation. A vector is containing both the lindan-resistance gene as a selectable marker, and GUS gene as a screenable marker. By using hypocotyl explant and vertical culture on B5 medium supplemented with 1 mg/1 kinetin- and 2,4-D 2 mg/1 and subcultured every 3-4 weeks, we can recommend to get a good and much callus from bean. This will help in introducing foreign DNA into callus cells. One strain of Agrobacterium carrying plasmid as vector for introducing foreign DNA into plant cells was used. At different concentrations of lindan; 3, 4 and 4.5 mg/I, the transformed Maxidor callus survived and grew over a period of 6 month and subcultured every 3-4 weeks, but the control callus died. Callus were assayed for GUS activity to confirm the expression of the GUS gene using the histochemical assay test. The GUS gene was also correctly expressed in callus cultures grown on 4mg/I lindan-selected medium, the typical blue colour in the histochemical assay using the X-gluc as substrate. But the control, non-transformed callus was not able to grow in the presence of lindan, neither showed a positive reaction in the in vitro assays.
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Molecular characterization of apricot (Prunus armeniaca L.) cultivars using cross species SSR amplification with peach primers
53-57.Views:332Apricot takes an important place in Hungarian fruit production. Considering morphological characteristics of apricots it was concluded that the genetics background of European cultivars is very limited. Molecular markers and their use for genotyping have revolutionized the identification of cultivars. In a classic apricot breeding program, it is important to be able to establish unique DNA profiles of selections to identify them unambiguously and to determine their genetic relationship. Presently SSR is far the most frequently performed technique for genetic diversity studies. In this study there were used peach and apricot primer pairs from four different sources in order to examine microsatellite polymorphism among cultivars and investigate relationships among them. The possibility of cross species amplification among different Prunus species using SSR primers allowed us to use primers developed in peach to study genetic diversity in apricot. In this work, 90% of the primers used were able to amplify SSRs in apricot and more than half of them were polymorphic. With the 10 primer pairs utilized were proven to be sufficient to set unique fingerprint for several cultivars studied. The obtained dendrogram classified of the 45 cultivars included in this study into two major groups and several subgroups.
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Callus induction on standard type Cymbidium cultivars
108-110.Views:311Tissue cultured Cymbidium PLBs (protocormlike body) were used as starting material to induce embryogenic callus which could serve as objects of genetic transformation. We obtained callus using two methods. The first method was culturing the PLB segments for one month in liquid MS medium in the presence of 0.5 mg/1 benzyladenine and 0.05 mg/1 naphtylacetic acid followed by cultivation on the same composition solid medium with 0.5 g/l activated charcoal for an additional month. Callus formation was observed on 30% of the explants. The second way was to propagate the PLB segments on solid MS medium supplemented with 1 mg/1 thidiazuron. In these cultures we also observed callus formation on 20% of the explants.
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Down-regulation of ethylene production in carnation (Dianthus Caryphyllus L.) by an apple derived ACC-cDNA
101-104.Views:215Transgenic carnations were produced with an apple derived antisense ACC-synthase cDNA. Transgenic carnation regenerants were potted in glasshouse. All transformed plants showed normal growth and were true-to-type. Ethylene production — measured at full opening stage — lowered by 30-60 %, no plant with 100 % decrease was identified. The vase-life has been observed for 5 years. 38 % of the transformant carnations showed a higher a relative value in days by more than 2 days to 6 days. Twenty six plants were found exhibiting the most marked alterations in the tested trait. In these plants ethylene production decreased by 37-67 %, they have longer vase-life (by 4 days or more). Since the fragrance variety 'Bíbor' was the plant material for genetic modification of vase-life, this trait has been conserved after transformation in spite of the fact that the position of transgene integration cannot be directed.
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Preliminary characterization of the self-incompatibility genotypes of European plum (Prunus domestica L.) cultivars
23-26.Views:420European plum is an important fruit crop with complex, hexaploid genome of unknown origin. The characterization of the selfincompatibility (S) locus of 16 European plum cultivars was carried out using the PaConsI-F primer in combination with the EM-PC1consRD primer for the first intron and the EM-PC2consFD and EM-PC3consRD primers for the second intron amplification. Altogether, 18 different alleles were scored indicating high genetic diversity. These alleles were labelled using alphabetical codes from SA to SS. We identified 5 different alleles in 9 cultivars, 4 alleles in 5 cultivars, while 3 alleles were shown in two of the assayed cultivars. A total of 16 different S-genotypes were assigned, and discrimination of all plum cultivars was successful based on their unique S-genotypes. However, further research is required to reliably identify the S-alleles based on their DNA sequence and clarify complete S-genotypes.
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Microsatellite based identification of grapevine cultivars traditional in Hungary and in the Carpathian Basin
71-73.Views:242Grapevine cultivars and clones traditional in Hungary and in the Carpathian Basin are maintained in the genetic collection of the Corvinus University of Budapest. Mostly ancient varieties and clones were genotyped using microsatellite loci. The investigated 6 loci were sufficient to distinguish all cultivars. Some clones could also be separated based mostly on the variable VVS2 loci. For 17 out of the 31 investigated cultivars this is the first report on characterization of the polymorphism of the allele lengths by microsatellite markers on loci: VVS2, VVMD7, VVMD27, vrZAG62.
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Factors affecting apricot fruit antioxidant capacity and mineral element contents
95-99.Views:423Several epidemiological studies revealed that the consumption of antioxidant compounds and the risk of atherosclerosis, increased blood pressure or cancer are inversely proportional. The individual amounts and relative proportions of macro- and micro elements present in food is also of great consideration since these are involved in a wide range of physiological processes including the influence of the redox homeostasis. The antioxidant power and mineral nutrient content of fruits might be affected by several factors including genotype, ripening stage, year-effects or a wide range of environmental conditions. This study was carried out to survey the antioxidant power and mineral element content in fresh fruits of apricot and analyse some genetic and environmental factors that may have important contribution to the inner content of apricot fruits. In addition, the influencing effect of the extraction procedure used for antioxidant analyses was also tested. Our analyses indicate that a considerable fraction of antioxidant capacity is attributable to the hydrophilic antioxidants. The genetic background has crucial importance in determining apricot fruit antioxidant capacity and mineral nutrient content; however, the growing season and the ripening time of fruits may have also important effects.
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Genotyping Hungarian apricot cultivars for self-(in)compatibility by isoelectric focusing and PCR analysis
69-72.Views:355Self-incompatibility (SI) in flowering plants is a widespread genetic system that promotes out-crossing. In Prunus species the SI is a gametophytic trait, which is controlled by a single multiallelic locus, termed S-locus. S-alleles codify stylar glycoproteins with ribonuclease activity (S-RNases). Our objective was to assess the S-genotype of some Hungarian apricot varieties by isoelectric focusing of stylar RNases as well as by PCR analysis using cherry consensus primers. Consensus primers amplified one or two bands of various sizes. Primers amplifying the 1st intron gained fragments the size of which ranged from 250 to 500 bp; while those amplifying the 2nd intron resulted in fragments of 800-2000 by length. Our data demonstrated that the first intron of the apricot S-RNase gene is shorter than the second one, which coincides with the structure of cherry S-RNase alleles. `Hargrand' (S1S2) and `Harcoe (S1S4) possessed one common S-RNase isoenzyme. Hungarian 'Orias' apricot cultivars showed different bands compared to the previous cultivars, but they shared completely identical patterns confirming that they possess the same S-genotype. 'Bergeron', `Harmat' and 'Korai zamatos' are characterised by an evidently distinct S-RNase pattern. The self-compatible cultivar (`Bergeron') had one allele, which suggests its correspondence to the Sc. Primers for the 2nd intron was unsuccessful in gaining fragments, which indicates that the 2nd intron in the Sc allele is too long to get any amplification. On the basis of our data, identities and differences were revealed in the S-allele constitution of some economically important Hungarian apricot cultivars at protein and DNA levels.
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The effects of ACS (1-aminocyclopropane-l-carboxylate synthase) gene down regulation on ethylene production and fruit softening in transgenic apple
65-70.Views:220A detailed examination of the production of ethylene and other ripening parameters during storage period has been undertaken in transgenic apple fruits, where the ethylene biosynthesis was inhibited by antisense ACS (l-aminocyclopropane-l-carboxylate synthase) gene. Data indicate down regulation of ethylene production, softening and spoilage in some transgenic lines. In some cases ethylene production was inhibited for over 90 percent, considerable reduction of softening and spoilage was observed probably due to the reduced activity of cell wall degradable enzymes. ACS activity was also monitored during ripening. The fruits of the best transgenic lines could be stored for minimum 4-5 months longer under 5 °C cold room storage conditions and one month longer at normal room temperature. This molecular approach can provide an alternative way to replace the commonly used and costly atmospheric storage of fruits.
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Analyses of Hungarian sour cherry germplasm with simple sequence repeat markers
27-31.Views:932Twenty-four sour cherry cultivars (genotypes), belonging to four cultivar groups were fingerprinted using microsatellite markers. All genotypes have been arisen from the Carpathian basin, which could be secondary gene centre of sour cherry, since its progenitor species, ground cherry and sweet cherry overlap here. Five SSR primer pairs, earlier used for fingerprinting Turkish sour cherry germplasm were tested. None of the five primer pairs showed any polymorphism within the cultivar groups. The primer pairs were able to distinguish between the cultivar groups. The Oblacsinszka and the Cigánymeggy cultivar groups were the most difficult to separate, while the Pándy cultivar group was the most distinguishable.
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Luminescence variations in cucumber (Cucumis sativus L.) leaves derived from different regeneration systems
50-52.Views:239Plants obtained from in vitro culture can show increased susceptibility to environmental stress conditions. In the process of their adaptation to natural conditions it requires monitoring of their physiological state. The methods used to check this phenomenon should estimate quickly and exactly the tolerance to suboptimal environmental factors. Such requirements are satisfied by the methods of measuring chlorophyll luminescence in vivo, e.g. fluorescence induction and delayed luminescence. The objects of our studies were cucumber plants regenerated from cultures of callus and embryogenic cell suspension, as well as the plants obtained from seeds. The plants derived from in vitro cultures displayed a poor physiological condition at the early phase of adaptation characterised by higher susceptibility both to stress caused by increased density of the light flux and low temperature (4 °C) in comparison with the plants obtained from seeds.
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Determination of the cold tolerance off sour cherry cultivars with frost treatments in climatic chamber
49-54.Views:563Nowadays, sour cherry buds can be seriously damaged by spring and winter frosts. Unlike other fruit species threatened by high frost damage, sour cherry cultivars have not been assessed for frost tolerance. The aim of .our survey was to establish the relative cold tolerance of the Hungarian cultivars after treatment in a climatic chamber, and to optimize the methodology formerly elaborated for the frost treatment of apricot. Fourteen cultivars of Hungarian sour cherry (Prunus cerasus) were used in the experiments, which spanned the winters of 2005/2006 and 2006/2007. Our data were used to rank cultivars in two groups according to their levels of cold resistance. We also recommend critical temperatures and treatment times for the testing of sour cherry cultivar resistance to cold in climatic chambers.
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General defense system in the plant kingdom III.
45-54.Views:283Our observations regarding the symptoms not fitting into, significantly differing from the hypersensitive defense system, which we noticed during the judgment of several plant species, symptoms provoked on several million plants have constituted a unified entity. They have provided evidence for the existence of a different plant defense system. We called this so far unknown basic response of plants to biotic effects as general defense system. This system defends them from the attack of numerous microbe species in the environment.
The evolutionary intermediate phase between the general and the specific, the two defense systems is the susceptible host—pathogen relation. The vertical resistance system of plants escaping from the susceptible host—pathogen relation, based on specific hypersensitive reaction also suggested the existence of a more original, general defense system and the susceptible host—pathogen relation developed as a result of the collapse of that system.
The evolutionary relation of the two defense systems is proved by the only recessive inheritance of the older general defense system and in the majority of cases dominant hereditary course of the specific defense system. In our experiences, the modifying genes of the recessive general defense system, in most cases, are behind the specific defense systems, which are known to have monogenic dominant hereditary course and react with hypersensitive tissue destruction. This seemingly striking genetic fact is explained by the following: the general defense system less dependent on environmental effects regulates much faster pathophysiological reaction than the specific resistance genes strongly dependant on environmental effects coding dominant hypersensitive reaction.
The general and specific defense reactions, the processes excluding the microbes attacking plants with compacting of cell growth and tissue destruction, which mean two opposite strategies, building on and regulating each other constitute the entity of resistance to plant disease.
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Review of the molecular background of self-incompatibility in rosaceous fruit trees
7-18.Views:394This review gives a presentation of the gametophytic self-incompatibility system in the roscaeous fruit trees. Studies to discover the pistil (S-ribonucleases) and pollen-part components (F-box molecules) are summarized and models for the self-incompatibility reactions as well as their molecular background are discussed. We describe how mutations within the S-RNase or F-box genes can contribute to the transition from self-incompatibility to the self-compatible phenotype in many fruit tree crops. The current state of the arts is compared to the information obtained in other plant species possessing similar incompatibility system.
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Evaluation of different sources of resistance for breeding powdery mildew resistant grapevine varieties
35-40.Views:265The appearance of resistance to powdery mildew was investigated on seedling progenies of interspecific crossings. Hybrids of American and East-Asian wild species and one Vitis vinifera variety (Janjal kara) were used as source of resistance in the crossings. The resistance of foliage and of berries were tested in the field at the time of vintage. The degree of resistance was ranked into six classes.
In American and Asian hybrid derivatives there were few highly resistant progenies, but in the hybrids of Janjal kara variety more than 90 % of the progenies proved to be highly resistant to powdery mildew. According to the distribution of classes of resistance in the populations the resistance of American and Asian sources are mostly influenced by additive gene affects. The resistance of Janjal kara seemed to be mono or digenic.
No close correlation was found between the powdery mildew resistance of the foliage and that of the berries.
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Transmethylation and the general defense reaction of plants
35-40.Views:280Plant breeding for resistance, namely building specific resistance genes into cultivated plants to ensure resistance against certain pathogen species, is a several-decade-long practice. While looking for purposes of failures appearing during the cultivation of varieties created in this way, a plant feature that ensures non-specific reactions against effects which evoke biotic stress attracted our attention. We named this plant defense form the general defense reaction. The general defense reaction is a fundamental attribute of the plant kingdom, fulfils the role of plant immune system and manifests itself in cell enlargement and cell division. Plants with a high level general defense reaction endure abiotic stresses as well.
In studying the biochemical background of the interaction of the general defense reaction and transmethylation, we found that transmethylation has important role in warding off both biotic and abiotic stresses. According to our observations, plants possessing high level general defense system are suitable for thorough examination of the process and plant physiological role of transmethylation. Biochemical studies also strengthened our observation, which has been taken on the basis of phenotype, that the general defense system can not be ignored during future plant breeding.
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In vitro multiplication and hardening of grapevine plants in aeriated media
15-18.Views:322In vitro cultures have widely been used in horticulture for rapid multiplication of new varieties and clones as well as to produce pathogen-free stock material. To improve efficient hardening and transfer in vitro grown grapevine plants were multiplied by cutting them into single-node internodes with the whole leaf. Microcuttings including the shoot tips were rooted in granulated perlite moisted with tapwater under sterile conditions. After 2-3 weeks the rooted microcuttings were supplied by nutrients and hardened by gradual opening and finally by complete removal of the lids of jars or plastic boxes used for growth. Using this method microcuttings of Vitis vinifera cvs. „Chardonnay", „Cabernet franc", „Riesling" and „Sauvignon blanc" and the rootstock varieties Vitis riparia x Vitis cinerea cv. „Barrier" and Vitis berlandieri x Vitis rupestris cv. „Richter 110" formed new roots and shoots and 100% of the tested plants survived the acclimatization procedure. Similar results were obtained when perlite was replaced with rockwool-, or pit-pot blocks. This method may highly increase the efficiency of producing pathogen-free propagating material and new transgenic lines.
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Resistance Gene Analogs (RGA) as a tool in fruit tree's breeding
7-15.Views:312Breeding for pest and disease resistance comes as a major objective behind the fruit traits. To increase the effectiveness of fruit resistance breeding application of the Marker Assisted Selection ( MAS) is advantageous. For generating molecular markers which enable the following of interesting traits basically two methods are available: targeted marker design based on conservative region of already known Resistance ( R) gene sequences or randomly generated markers. The creation and the application of these homology based markers are the object of this review in the main temperate zone fruit species.
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Self-(in)compatibility in sour cherry (Prunus cerasus L.). A minireview
117-120.Views:304Sour cherry (Prunus cerasus L.) is an allotetraploid species derived from hybridisation of the diploid sweet cherry (P avium L.) and the tetraploid ground cherry (P. fruticosa Pall.). Although numerous self-incompatible cultivars exist, the most sour cherry cultivars are self-compatible, which might be due to their tetraploid nature. This review is dedicated to show the limited information on the genetics of self-incompatibility in sour cherry accumulated during the last five years. Two different hypotheses (genomic arrangement of the alleles or the accumulation of non-functional S-haplotypes) are discussed. Heteroallelic sour cherry pollen was shown to be self-incompatible, which is counter to the Solanaceae where heteroallelic pollen frequently self-compatible due to a kind of competitive interaction between the two different alleles. This review highlights some inconsistencies in the hope that clarification will be achieved in the near future.
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Large variations in antioxidant capacity and contents of Hungarian sour and sweet cherry cultivars
25-28.Views:809Sour cherry (Prunus cerasus L.) cultivars grown in Hungary are of local origin while most sweet cherry cultivars in Hungary are introduced from other countries.A great phenotypic variability is displayed by both species. In the present study, we analyzed 10 sour and 9 sweet cherry cultivars for their antioxidant capacity, total polyphenolics (TPC) and total anthocyanin (TMAC) contents. In general, sour cherries showed higher levels of antioxidant capacity, TPC and TMAC. The anthocyanin contents varied from 0.16 to 6.85 and 1.41 to 127.56 mg/100 g for sweet and sour cherries, respectively. However, TMAC generally seems to have a limited influence on the antioxidant capacity of cherries.An amarelletype sour cherry, ‘Pipacs 1’ showed the highest antioxidant capacity (21.21mmolAA/l) and TPC (44.07mgGA/l) in contrast to its lowanthocyanin content. The detected diversity presents a choice that can satisfy different consumer preferences, and meet specific nutritional requirements.
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Pollen tube growth in sweet cherry (Prunus avium L.) styles following fully compatible, half compatible and incompatible pollinations
63-68.Views:215In vivo as well as in vitro pollen tube growth studies along the style were performed, each with two pairs of sweet cherry cultivar combinations by means of fluorescence microscopy. In vivo studies showed that the percentage of pollen tubes penetrating the middle and basal section of the style was higher in the fully compatible 'Margit' x 'Alex' combination than in the half compatible `Germersdorfi 3' x `Alex' cross. The year effect was significant at P=0.] probability level. All pollen tubes in vitro stopped at the upper third of the style in the incompatible 'Vera' x 'Van' cross, whereas in the half compatible 'Alex' x 'Van' 50% of the pollen tubes penetrated to the lower third of the style. By in vitro fluorescence microscopy, it was possible to distinguish half compatible combinations from incompatible ones. Results obtained by in vivo technique only were much ambiguous.
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Recent findings of the tree fruit self-incompatibility studies
7-15.Views:373This review endeavours to collect all recent and substantial contributions to the quickly deepening fields of tree fruit self-incompatibility studies and hence updating previously published reviews. Studies carried out to discover the molecular basis of gametophytic self-incompatibility are summarized and a newly described model for the solanaceous plants is also outlined. We describe recent findings in all economically important fruit tree crops involving apple, European pear, sweet and sour cherries, almond, Japanese plum, sloe, Japanese apricot, European apricot and peach. Additional DNA sequences are now available for both the pistil and pollen component genes in several species and their molecular, evolutionary or economic implications are discussed in the light of the fruit setting behaviour.
